We propose to continue studies on the molecular biology and life cycle of the human papillomaviruses (HPVs). There are at least 15 distinct HPVs. Very little is known about them because they do not grow in cell culture. They differ from conventional tumor viruses in several respects, their replication is tied to cell differentiation, and they have enormous medical significance. HPVs cause considerable debilitating and disfiguring human disease. In addition, HPV genetic information has been found in human urogenital cancers, in squamous cell carcinomas of patients with epidermodysplasia verruciformis, and in cervical dysplasia. In this proposal we will: (1) complete the DNA sequence of the cloned HPV-3 genome and sequence the cloned HPV-5 genome; (2) synthesize peptides targeted to specific domains of putative HPV-coded proteins (based on the DNA sequence) and prepare antibodies to each synthetic peptide; (3) establish experimental cell systems to study HPV gene expression; (4) analyze the physical state of the HPV genome (copy number, integrity of the viral genome, and whether integrated or episomal), characterize HPV transcripts and mRNAs, and identify HPV-coded proteins in experimentally transformed cells; (5) analyze HPV-coded RNAs and proteins in cells and tissues naturally harboring HPVs, e.g. cultured human laryngeal papilloma cells, condylomas, plantar warts, sequential thin sections of cervical dysplasia biopsies; and (6) explore the construction of HPV "shuttle vectors" that could replicate as episomes in E. coli and mammalian cells, especially human cells, and that could be used to introduce cloned genes into mammalian cells for functional analysis.